title: Screening protein--single stranded RNA complexes by NMR spectroscopy for structure determination.
abstract: In the past few years, RNA molecules have been revealed to be at the center of numerous biological processes. Long considered as passive molecules transferring genetic information from DNA to proteins, it is now well established that RNA molecules play important regulatory roles. Associated with that, the number of identified RNA binding proteins (RBPs) has increased considerably and mutations in RNA molecules or RBP have been shown to cause various diseases, such as cancers. It is therefore crucial to understand at the molecular level how these proteins specifically recognise their RNA targets in order to design new generation drug therapies targeting protein-RNA complexes. Nuclear magnetic resonance (NMR) is a particularly well-suited technique to study such protein-RNA complexes at the atomic level and can provide valuable information for new drug discovery programs. In this article, we describe the NMR strategy that we and other laboratories use for screening optimal conditions necessary for structural studies of protein-single stranded RNA complexes, using two proteins, Sam68 and T-STAR, as examples.
authors.affiliation: Department of Biochemistry, Henry Wellcome Laboratories of Structural Biology, University of Leicester, UK.
authors.forename: Jaelle N
authors.initials: JN
authors.lastname: Foot
authors.affiliation: Department of Biochemistry, Henry Wellcome Laboratories of Structural Biology, University of Leicester, UK.
authors.forename: Mikael
authors.initials: M
authors.lastname: Feracci
authors.affiliation: Department of Biochemistry, Henry Wellcome Laboratories of Structural Biology, University of Leicester, UK. Electronic address: cyril.dominguez@leicester.ac.uk.
authors.forename: Cyril
authors.initials: C
authors.lastname: Dominguez
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000235:genetics; D000595:Amino Acid Sequence; D052157:Aptamers, Nucleotide / Q000138:chemical synthesis / Q000737:chemistry*; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D052157:Aptamers, Nucleotide; D004268:DNA-Binding Proteins; C489559:KHDRBS1 protein, human; C118777:KHDRBS3 protein, human; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins
keywords: NMR spectroscopy; Protein–RNA complex; STAR proteins; Sam68; T-STAR